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Background  The enteroaggregative  E. coli  heat-stable enterotoxin 1 (EAST1) encoded by  astA  gene has been found in enteropathogenic  E. coli  (EPEC) strains. However, it is not sufficient to simply probe strains with an  astA  gene probe due to the existence of  astA  mutants (type 1 and type 2 SHEAST) and EAST1 variants (EAST1 v1-4). In this study, 222 EPEC (70 typical and 152 atypical) isolates were tested for the presence of the  astA  gene sequence by PCR and sequencing.    Results  The  astA  gene was amplified from 54 strains, 11 typical and 43 atypical. Sequence analysis of the PCR products showed that 25 strains, 7 typical and 18 atypical, had an intact  astA  gene. A subgroup of 7 atypical strains had a variant type of the  astA  gene sequence, with four non-synonymous nucleotide substitutions. The remaining 22 strains had mutated  astA  gene with nucleotide deletions or substitutions in the first 8 codons. The RT-PCR results showed that the  astA  gene was transcribed only by the strains carrying either the intact or the variant type of the  astA  gene sequence. Southern blot analysis indicated that  astA  is located in EAF plasmid in typical strains, and in plasmids of similar size in atypical strains. Strains carrying intact  astA  genes were more frequently found in diarrheic children than in non-diarrheic children (p < 0.05).    Conclusion  In conclusion, our data suggest that the presence of an intact  astA  gene may represent an additional virulence determinant in both EPEC groups.

bmc microbiology

Detection and genetic analysis of the enteroaggregative Escherichia coli heat-stable enterotoxin (EAST1) gene in clinical isolates of enteropathogenic Escherichia coli(EPEC) strains